JP6266347B2 - Marker peptide for determining risk of developing metabolic syndrome and use thereof - Google Patents

Description

  The present invention relates to a marker peptide for determining risk of suffering from metabolic syndrome and use thereof.

  At present, the number of patients with metabolic syndrome is increasing in Japan, and the total number of affected persons is estimated to be 20 million when combined with the metabolic syndrome reserve army. Early detection and prevention are important because metabolic syndrome can cause arteriosclerosis in severe cases, leading to myocardial or cerebral infarction. In April 2008, specific screening (Metabo screening) became mandatory for early detection. Metabolic syndrome prevention measures include food intake, and functional foods including foods for specified health use that obesity and / or prevent metabolic syndrome are on the market.

  On the other hand, various marker substances that are contained in living bodies and serve as indices for disease diagnosis have been reported. For example, Non-Patent Document 1 describes that P-B peptide, which is a proline-rich peptide in saliva, is significantly reduced in children with type 1 diabetes. Non-Patent Document 2 describes that the polymorphism of the GIPR gene has a correlation with the blood glucose level after 2 hours in the glucose tolerance test.

Mol Cell Proteomics. 2010 Oct: 9 (10): 2099-108 Nature Genet. 2010 Feb: 42 (2): 142-148

  However, the metabolic syndrome prevention effect of functional foods is difficult to see and there is a problem that it is difficult for consumers to maintain motivation to continue using. No marker substance that can be used as an indicator of metabolic syndrome has been found so far.

  An object of the present invention is to provide a marker substance that can accurately determine the risk of suffering from metabolic syndrome in a living body.

  As a result of intensive studies, the present inventors have found that a part of the amino acid sequences of the P-B peptide and GIPR are useful as marker peptides for determining the risk of suffering from metabolic syndrome, and biological samples such as saliva It was found that the risk of suffering from metabolic syndrome can be determined by determining the amount of the marker peptide. The present invention is based on such knowledge.

The present invention provides the following [1] to [12].
[1] A marker peptide for determining risk of developing metabolic syndrome, selected from the group consisting of the following (A) to (J).
(A) a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1;
(B) For determining the risk of developing metabolic syndrome, including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker;
(C) a polypeptide comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining risk of developing metabolic syndrome;
(D) In the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added, substituted or inserted, and contain 10 or more consecutive amino acid residues, and risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(E) a polypeptide comprising 10 or more consecutive amino acid residues in an amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome;
(F) a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2;
(G) For determining the risk of developing metabolic syndrome, including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2. A polypeptide that can be used as a marker;
(H) a polypeptide comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2;
(I) In the amino acid sequence of SEQ ID NO: 2, one or several amino acids are deleted, added, substituted or inserted, and contain 10 or more consecutive amino acid residues, and risk of suffering from metabolic syndrome A polypeptide that can be used as a marker; and (J) comprising 10 or more consecutive amino acid residues of an amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 2, and determining risk of suffering from metabolic syndrome [2] An antibody or aptamer that binds to the marker peptide for determining metabolic syndrome morbidity risk described in [1].
[3] A microarray in which an antibody or aptamer that binds to the metabolic syndrome disease risk determination marker peptide according to [1] above is immobilized on a carrier.
[4] A method for determining metabolic syndrome morbidity risk, comprising measuring the amount or presence of the marker syndrome for metabolic syndrome morbidity determination according to [1] above in a biological sample collected from a subject.
[5] When the amount of the metabolic syndrome morbidity risk determination marker peptide according to [1] above in the biological sample collected from the subject is higher than the amount in the biological sample collected from healthy subjects, The determination method according to the above [4], wherein the risk of causing syndrome is high.
[6] The amount of the metabolic syndrome morbidity risk determination marker peptide according to [1] above in a biological sample collected from a subject is the same or higher than the amount in a biological sample collected from a patient with metabolic syndrome The determination method according to [4] above, wherein the risk of suffering from metabolic syndrome is determined to be high.
[7] The measurement of the amount of the marker syndrome for determining metabolic syndrome risk described in [1] above, mass spectrometry, immunoassay using the antibody or aptamer described in [2], and [3] The determination method according to any one of [4] to [6], wherein the determination is performed by a method selected from immunoassays using the microarray.
[8] The determination method according to [4] above, wherein multivariate analysis is performed using a combination of two or more marker peptides for risk determination of metabolic syndrome as a variable.
[9] The determination method according to [8], wherein the multivariate analysis is logistic regression analysis.
[10] The determination method according to any one of [4] to [9], wherein the biological sample is saliva.
[11] A kit for determining the risk of developing metabolic syndrome, comprising the antibody or aptamer of [2] above or the microarray of [3] above.
[12] The kit according to [11], further including a gum.

The present invention can also be applied to diagnosis of metabolic syndrome. Examples of methods for diagnosing metabolic syndrome include the following modes.
[13] A marker peptide for diagnosis of metabolic syndrome selected from the group consisting of the above (A) to (J).
[14] An antibody or aptamer that binds to the metabolic syndrome diagnostic marker peptide of [13] above.
[15] A microarray in which an antibody or aptamer that binds to the metabolic syndrome diagnostic marker peptide of [13] above is immobilized on a carrier.
[16] A method for diagnosing metabolic syndrome, comprising measuring the amount or presence of the marker peptide for diagnosing metabolic syndrome according to [13] above in a biological sample collected from a subject.
[17] The diagnostic method according to [16] above, wherein the subject is a human.
[18] When the amount of the metabolic syndrome diagnostic marker peptide according to [13] above in the biological sample collected from the subject is higher than the amount in the biological sample collected from a healthy subject, the metabolic syndrome The diagnostic method according to [16] or [17] above, wherein it is determined that the patient is affected.
[19] When the amount of the metabolic syndrome diagnostic marker peptide according to the above [13] in the biological sample collected from the subject is the same or higher than the amount in the biological sample collected from a metabolic syndrome patient, The diagnostic method according to the above [16] or [17], wherein it is determined that the subject has a metabolic syndrome.
[20] The amount of the metabolic syndrome diagnostic marker peptide described in [13] is measured by mass spectrometry, an immunoassay using the antibody or aptamer described in [14], and the microarray described in [15] The diagnostic method according to any one of [16] to [19], which is performed by a method selected from immunoassays using
[21] The diagnostic method according to [16] or [17] above, wherein multivariate analysis is performed using a combination of two or more metabolic syndrome diagnostic marker peptides as variables.
[22] The diagnostic method according to [21], wherein the multivariate analysis is logistic regression analysis.
[23] The diagnostic method according to any one of [16] to [22], wherein the biological sample is saliva.
[24] A metabolic syndrome diagnostic kit comprising the antibody or aptamer of [14] above or the microarray of [15] above.
[25] The kit according to [24], further including a gum.

  According to the present invention, it is possible to accurately determine the risk of suffering from metabolic syndrome. If the risk of morbidity of metabolic syndrome is determined prior to diagnosis by a doctor using the present invention, preventive measures for metabolic syndrome can be taken early, and morbidity of metabolic syndrome can be prevented in advance.

FIG. 1 is a diagram showing the peak intensity of ^PB peptide fragment (1) ^27-54 in each subject group. FIG. 2 is a diagram showing peak intensities of GIPR peptide fragments in each subject group. FIG. 3 is a diagram showing the peak intensity of the ^PB peptide fragment (2) ^23-54 in each subject group. FIG. 4 is a diagram showing the peak intensity of ^PB peptide fragment (3) ^55-79 in each subject group. FIG. 5 is a diagram showing the peak intensity of the ^PB peptide fragment (4) ^23-79 in each subject group. FIG. 6 is a diagram showing the peak intensity of the ^PB peptide fragment (5) ^23-35 in each subject group.

  The present invention relates to risk determination of metabolic syndrome. In the present invention, the determination of the risk of suffering from metabolic syndrome is the determination of whether or not a subject is suffering from metabolic syndrome, whether it has already been cured, or whether there is a possibility of suffering in the future (evaluation, discrimination, discrimination, estimation) ) Or a distinction (classification) of whether or not the subject has a risk of suffering from metabolic syndrome. Determining the risk of morbidity means determining whether a subject has developed metabolic syndrome, has already been cured, and is likely to develop in the future. The accuracy of determination of morbidity risk is such that the risk of morbidity of metabolic syndrome can be correctly determined in a statistically significant proportion of subjects, for example, 50% or more, 60% or more, 70% or more, for example. 80% or more, 85% or more, or 90% or more of the subjects can correctly determine the risk of suffering from metabolic syndrome. The determination method of the present invention is useful as a preliminary determination method before diagnosis by a doctor.

  In the present invention, metabolic syndrome refers to a state in which hyperglycemia, hypertension or hyperlipidemia is caused by visceral fat obesity. According to the standard (2005) defined by the Japanese Society of Obesity for Japanese, this means that any one or more of items a) to d) shown in the examples are applicable.

  The subject in the present invention is usually an animal, preferably a human or a laboratory animal (mouse, rat, guinea pig, hamster, rabbit, etc.), more preferably a human.

(1) Marker peptide of this invention The marker peptide for metabolic syndrome morbidity risk determination of this invention is 1 or more selected from the group which consists of said (A)-(J).

  When determining the risk of developing metabolic syndrome using the marker peptide for determining the risk of developing metabolic syndrome of the present invention, one type selected from (A) to (J) may be used, but higher accuracy is achieved by combining two or more types. Judgment is possible. In the case of two or more types of combinations, a combination of one or more types of peptides selected from the group consisting of (A) to (E) and one or more types of peptides selected from the group consisting of (F) to (J) Preferably there is. Preferred examples of combinations of two or more types are listed in item (4).

(1-1) About (A) to (C) In (A) to (C), the amino acid sequence of SEQ ID NO: 1 is a human P-B peptide (Sub-grilled grand-and-regulated protein 3B, Proline-rich peptide P- B, an amino acid sequence encoding Proline-rich protein 3) (79 amino acids in total length). The P-B peptide is a peptide belonging to the proline rich protein (PRP) family that is rich in proline (P).

  In (A) to (C), 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1 are 11 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. Is more preferable, it is more preferably 12 or more consecutive amino acid residues, and even more preferably 13 or more consecutive amino acid residues. The upper limit is not particularly limited, and may be the full length (79 residues) of the amino acid sequence of SEQ ID NO: 1 or more. For example, 100 or less, 90 or less, 80 or less, 70 or less, 60 Or less.

  In (A) to (C), in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54. Part, at least part of positions 23 to 54, at least part of positions 55 to 79, at least part of positions 23 to 79, or at least part of positions 23 to 35. 10 or more amino acid residues are 10 or more consecutive amino acid residues in amino acid residue positions 27 to 54 of SEQ ID NO: 1, and 10 or more consecutive amino acids in positions 23 to 54 Residue, 10 or more consecutive amino acid residues in positions 55 to 79, 10 or more consecutive amino acid residues in positions 23 to 79, or 10 consecutive positions in positions 23 to 35 It is more preferable that the above amino acid residues are included. 10 or more amino acid residues are amino acid residues Nos. 27 to 54 in SEQ ID NO: 1, amino acid sequences Nos. 23 to 54, amino acids Nos. 55 to 79, and Nos. 23 to 79. More preferred is an amino acid sequence, an amino acid sequence at positions 23 to 35, or an amino acid sequence of SEQ ID NO: 1.

  (A) may be any polypeptide containing 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. A polypeptide consisting of

  (B) includes an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and has a risk of suffering from metabolic syndrome Any polypeptide that can be used as a marker for determination may be used, and one or several amino acids are deleted, added, substituted, or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. A polypeptide that consists of a sequence and can be used as a marker for risk assessment of metabolic syndrome is preferable.

  (C) is a polypeptide that includes an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and can be used as a marker for determining the risk of developing metabolic syndrome. It is a polypeptide that can be used as a marker for risk assessment of metabolic syndrome, comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. It is preferable.

  In (B), the mutation of one or several amino acid residues may be present in one region in the amino acid sequence, but may be present in a plurality of different regions. The term “one or several” indicates a range that does not significantly impair the function or properties of the marker peptide. “One or several” means, for example, “one or three”, “one or two” or “one”.

  The positions of amino acid residues that are allowed to be mutated (deletion, addition, substitution or insertion) in the amino acid sequence are obvious to those skilled in the art. Specifically, those skilled in the art will 1) amino acid sequences of a plurality of proteins having the same type of activity (for example, the amino acid sequence represented by SEQ ID NO: 1 and the amino acid sequences of other marker syndromes for determining the risk of metabolic syndrome) 2) reveal the relatively conserved and relatively unconserved regions, then 3) from the relatively conserved and relatively unconserved regions Since it is possible to predict regions that can play an important role in function and regions that can not play an important role in function, it is possible to recognize the correlation between structure and function. Therefore, those skilled in the art can estimate the position of an amino acid residue that is allowed to be mutated in the amino acid sequence of the marker for determining risk of developing metabolic syndrome.

  In the case of including mutation due to amino acid residue substitution, the amino acid residue substitution may be a conservative substitution. As used herein, the term “conservative substitution” refers to the replacement of a given amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are well known in the art, such as amino acids having basic side chains (eg, lysine, arginine, histidine), amino acids having acidic side chains (eg, aspartic acid). , Glutamic acid), amino acids having uncharged polar side chains (eg, asparagine, glutamine, serine, threonine, tyrosine, cysteine), amino acids having nonpolar side chains (eg, glycine, alanine, valine, leucine, isoleucine, proline) , Phenylalanine, methionine, tryptophan), amino acids having a β-branched side chain (eg, threonine, valine, isoleucine), amino acids having aromatic side chains (eg, tyrosine, phenylalanine, tryptophan, histidine), hydroxyl groups (eg, Alcoholic and phenolic) Amino acids having a chain (eg, serine, threonine, tyrosine), and amino acids having aromatic side chains (e.g., cysteine, methionine) and the like. Preferably, the conservative substitution of amino acids is a substitution between aspartic acid and glutamic acid, a substitution between arginine and lysine and histidine, a substitution between tryptophan and phenylalanine, and between phenylalanine and valine. Or a substitution between leucine, isoleucine and alanine, and a substitution between glycine and alanine.

  In (C), the homology (for example, identity, similarity) to the amino acid sequence is 90% or more, preferably 95%, more preferably 98% or more, and further preferably 99% or more.

  Amino acid sequence homology (eg, identity, similarity) is determined by, for example, the algorithm BLAST (Pro. Natl. Acad. Sci. USA, 90, 5873 (1993)) by Karlin and Altschul or FASTA (Methods Enzymol., 183) by Pearson. 63 (1990)). Since programs called BLASTP and BLASTN have been developed based on the algorithm BLAST (see http://www.ncbi.nlm.nih.gov), these programs are used with default settings, and amino acid sequence homology is used. Sex may be calculated.

Examples of (A) include (A-1) to (A-33):
(A-1) a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1;
(A-2) It contains 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising a moiety;
(A-3) Consists of 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising a moiety;
(A-4) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 27 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-5) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 27 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-6) a polypeptide comprising amino acid residue numbers 27 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-7) a polypeptide comprising amino acid residues Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-8) It contains 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising a moiety;
(A-9) It consists of 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising a moiety;
(A-10) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-11) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue positions 23 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-12) a polypeptide comprising amino acid residues 23 to 54 in the amino acid sequence of SEQ ID NO: 1;
(A-13) a polypeptide comprising amino acid residues Nos. 23 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-14) It contains 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue positions 55 to 79 A polypeptide comprising a moiety;
(A-15) It consists of 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 55 to 79. A polypeptide comprising a moiety;
(A-16) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue positions 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-17) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue positions 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-18) a polypeptide comprising amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-19) a polypeptide comprising amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-20) including 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 79 A polypeptide comprising a moiety;
(A-21) It consists of 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 79. A polypeptide comprising a moiety;
(A-22) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-23) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-24) a polypeptide comprising amino acid residues Nos. 23 to 79 in the amino acid sequence of SEQ ID No. 1;
(A-25) a polypeptide consisting of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-26) comprising 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 35 A polypeptide comprising a moiety;
(A-27) Consisting of 10 or more amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue positions 23 to 35 A polypeptide comprising a moiety;
(A-28) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue positions 23 to 35 in the amino acid sequence of SEQ ID NO: 1;
(A-29) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 35 in the amino acid sequence of SEQ ID NO: 1;
(A-30) a polypeptide comprising amino acid residues 23 to 35 in the amino acid sequence of SEQ ID NO: 1;
(A-31) a polypeptide comprising amino acid residues Nos. 23 to 35 in the amino acid sequence of SEQ ID No. 1;
(A-32) a polypeptide comprising the amino acid sequence of SEQ ID NO: 1; and (A-33) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1.

Examples of (B) include (B-1) to (B-33):
(B-1) Risk of suffering from metabolic syndrome consisting of an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for determination;
(B-2) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(B-3) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(B-4) In 10 or more consecutive amino acid residues selected from amino acid residue Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(B-5) In 10 or more consecutive amino acid residues selected from amino acid residue Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(B-6) including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid residue Nos. 27 to 54 of the amino acid sequence of SEQ ID No. 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-7) It consists of an amino acid sequence in which amino acid residues Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1 are deleted, added, substituted or inserted, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-8) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(B-9) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(B-10) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 54 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(B-11) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 54 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(B-12) including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-13) Complicated with metabolic syndrome, comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for risk determination;
(B-14) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 55 to 79 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(B-15) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 55 to 79 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(B-16) In 10 or more consecutive amino acid residues selected from amino acid residue positions 55 to 79 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(B-17) In 10 or more consecutive amino acid residues selected from amino acid residue positions 55 to 79 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(B-18) including an amino acid sequence in which one or several amino acids are deleted, added, substituted, or inserted in the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-19) consisting of an amino acid sequence in which one or several amino acids are deleted, added, substituted, or inserted in the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-20) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue positions 23 to 79 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(B-21) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue positions 23 to 79 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(B-22) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 79 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(B-23) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 79 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(B-24) including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid residue Nos. 23 to 79 in the amino acid sequence of SEQ ID No. 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-25) An amino acid sequence of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1, consisting of an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-26) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 35 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(B-27) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 35 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(B-28) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 35 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(B-29) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 23 to 35 in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(B-30) including an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid residue Nos. 23 to 35 of the amino acid sequence of SEQ ID No. 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-31) consisting of an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(B-32) a polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome; and (B -33) A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome.

Examples of (C) include (C-1) to (C-33):
(C-1) a polypeptide comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, which can be used as a marker for risk assessment of metabolic syndrome;
(C-2) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(C-3) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 27 to 54 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(C-4) including an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue numbers 27 to 54 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-5) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1; A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-6) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-7) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid residues of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-8) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(C-9) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 54 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(C-10) comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residues 23 to 54 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-11) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-12) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid residue positions 23 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-13) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-14) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 55 to 79 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(C-15) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 55 to 79 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(C-16) comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue positions 55 to 79 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-17) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-18) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-19) A polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 55 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-20) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 79 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(C-21) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 79 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(C-22) including an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue positions 23 to 79 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-23) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-24) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid residue positions 23 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-25) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-26) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue numbers 23 to 35 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(C-27) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1, and the 10 or more amino acid residues include at least one of amino acid residue positions 23 to 35 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(C-28) comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue positions 23 to 35 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-29) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue positions 23 to 35 in the amino acid sequence of SEQ ID NO: 1, A polypeptide that can be used as a marker for risk assessment of syndrome;
(C-30) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-31) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-32) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining risk of developing metabolic syndrome; and (C-33) amino acid sequence of SEQ ID NO: 1 And a polypeptide that can be used as a marker for determining the risk of developing metabolic syndrome.

(1-2) (D) to (E) In (D) to (E), the amino acid sequence of SEQ ID NO: 1 is as described in item (1-1) above.

  In (D) to (E), 10 or more consecutive amino acid residues in the amino acid sequence having each mutation in the amino acid sequence of SEQ ID NO: 1 are 11 or more consecutive amino acid residues. It is more preferable that the amino acid residues are 12 or more consecutive amino acid residues, and it is more preferable that the amino acid residues are 13 or more consecutive amino acid residues. The upper limit is not particularly defined, and the amino acid sequence of SEQ ID NO: 1 may have a full length (79 residues) or more of the amino acid sequence having each mutation, but for example, 100 or less, 90 or less, 80 or less, 70 or less, or 60 or less.

  In (D), in the amino acid sequence of SEQ ID NO: 1, one or several amino acids are deleted, added, substituted or inserted, and contain 10 or more consecutive amino acid residues, and suffer from metabolic syndrome The polypeptide may be any polypeptide that can be used as a marker for risk determination. In the amino acid sequence of SEQ ID NO: 1, 10 or more consecutive amino acid sequences in which one or several amino acids are deleted, added, substituted, or inserted. A polypeptide that consists of amino acid residues and can be used as a marker for risk assessment of metabolic syndrome is preferable.

  (E) is a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome A polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome It is preferable that

  The mutation of one or several amino acid residues in (D) is the same as that described for (B) in item (1-1) above.

  The homology in (E) is the same as that described for (C) in item (1-1) above.

  As a peptide selected from the group consisting of (A) to (E), (A-2) to (A-31), (B-2) to (B-31) and (C-2) to (C-) Peptide selected from the group consisting of 31) is preferred, and (A-6), (A-7), (A-12), (A-13), (A-18), (A-19), (A -24), (A-25), (A-30) and a peptide selected from the group consisting of (A-31) are more preferred, (A-7), (A-13), (A-19), More preferred is a peptide selected from the group consisting of (A-25) and (A-31).

(1-3) About (F) to (H) In (F) to (H), the amino acid sequence of SEQ ID NO: 2 is human GIPR (gastric function inhibitory polypeptide receptor, glucose-dependent insulinotropic hormone receptor) , Which is an amino acid sequence encoding Gustic inhibitory polypeptide receptor and Glucose-dependent insulative polypeptide receptor (total length 466 amino acids).

  In (F) to (H), 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2 are 11 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2. And more preferably 12 or more consecutive amino acid residues. The upper limit is not particularly specified, and may be longer than the full length (466 residues) of the amino acid sequence of SEQ ID NO: 1. For example, 500 or less, 400 or less, 300 or less, 200 or less, 100 Or less, 90 or less, 80 or less, 70 or less, 60 or less, 50 or less, 40 or less, 30 or less, 25 or less, 20 or less, or 15 or less.

  In (F) to (H), in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275. It is preferable that 10 or more amino acid residues are 10 or more consecutive amino acid residues in amino acid residue numbers 264 to 275 of SEQ ID NO: 2. More preferably, 10 or more amino acid residues are the amino acid sequences of amino acid residues 264 to 275 of SEQ ID NO: 2, or the amino acid sequence of SEQ ID NO: 2.

  (F) may be any polypeptide containing 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2 A polypeptide consisting of

  (G) includes an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and has a risk of developing metabolic syndrome Any polypeptide that can be used as a marker for determination may be used, and one or several amino acids are deleted, added, substituted, or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2. A polypeptide that consists of a sequence and can be used as a marker for risk assessment of metabolic syndrome is preferable.

  (H) is a polypeptide comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for determining the risk of developing metabolic syndrome. It is a polypeptide that can be used as a marker for risk assessment of metabolic syndrome, comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 1. It is preferable.

  The mutation of one or several amino acid residues in (G) is the same as that described for (B) in item (1-1) above.

  The homology in (H) is the same as that described for (C) in item (1-1) above.

Examples of (F) include (F-1) to (F-9):
(F-1) a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2;
(F-2) It comprises 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275. A polypeptide comprising a moiety;
(F-3) It consists of 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275. A polypeptide comprising a moiety;
(F-4) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 in the amino acid sequence of SEQ ID NO: 2;
(F-5) a polypeptide comprising 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 in the amino acid sequence of SEQ ID NO: 2;
(F-6) a polypeptide comprising amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2;
(F-7) a polypeptide comprising amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2;
(F-8) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2; and (F-9) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2.

Examples of (G) include (G-1) to (G-9):
(G-1) It consists of an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and has a risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for determination;
(G-2) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted, and used as a marker for risk assessment of metabolic syndrome;
(G-3) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275 A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in a part of the amino acid sequence, and can be used as a marker for risk assessment of metabolic syndrome;
(G-4) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 in the amino acid sequence of SEQ ID NO: 2, one or several amino acids are deleted, added or substituted Or a polypeptide that includes an inserted amino acid sequence and can be used as a marker for determining the risk of developing metabolic syndrome;
(G-5) In 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 in the amino acid sequence of SEQ ID NO: 2, one or several amino acids are deleted, added or substituted Or a polypeptide comprising an inserted amino acid sequence, which can be used as a marker for risk assessment of metabolic syndrome;
(G-6) including the amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids are deleted, added, substituted or inserted, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(G-7) It consists of an amino acid sequence of amino acid residues 264 to 275 in the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids are deleted, added, substituted or inserted, and suffers from metabolic syndrome A polypeptide that can be used as a marker for risk determination;
(G-8) a polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for determining the risk of developing metabolic syndrome; and (G -9) A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO: 2, and can be used as a marker for determining risk of developing metabolic syndrome.

Examples of (H) include (H-1) to (H-9):
(H-1) a polypeptide comprising an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, which can be used as a marker for risk assessment of metabolic syndrome;
(H-2) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2 and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence comprising a portion, and being usable as a marker for risk assessment of metabolic syndrome;
(H-3) 10 or more consecutive amino acid residues in the amino acid sequence of SEQ ID NO: 2, and the 10 or more amino acid residues include at least one of amino acid residue numbers 264 to 275 A polypeptide comprising an amino acid sequence having a homology of 90% or more with an amino acid sequence containing a portion and usable as a marker for risk assessment of metabolic syndrome;
(H-4) including an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 of the amino acid sequence of SEQ ID NO: 2, A polypeptide that can be used as a marker for risk assessment of syndrome;
(H-5) consisting of an amino acid sequence having 90% or more homology with 10 or more consecutive amino acid residues selected from amino acid residue numbers 264 to 275 of the amino acid sequence of SEQ ID NO: 2; A polypeptide that can be used as a marker for risk assessment of syndrome;
(H-6) a polypeptide comprising an amino acid sequence having 90% or more homology with an amino acid residue at positions 264 to 275 of the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome ;
(H-7) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid residues of positions 264 to 275 of the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome ;
(H-8) a polypeptide comprising an amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 2 and usable as a marker for determining the risk of metabolic syndrome; and (H-9) amino acid sequence of SEQ ID NO: 2 And a polypeptide that can be used as a marker for determining the risk of developing metabolic syndrome.

(1-4) About (I) to (J) In (I) to (J), the amino acid sequence of SEQ ID NO: 2 is as described in item (1-3) above.

  In (I) to (J), 10 or more consecutive amino acid residues in the amino acid sequence having each mutation in the amino acid sequence of SEQ ID NO: 2 are 11 or more consecutive amino acid residues. The amino acid residue is more preferably 12 or more consecutive amino acid residues. The upper limit is not particularly specified, and the total length (466 residues) of the amino acid sequence having each mutation in the amino acid sequence of SEQ ID NO: 1 or may exceed this, but for example, 500 or less, 400 or less, 300 100 or less, 100 or less, 90 or less, 80 or less, 70 or less, 60 or less, 50 or less, 40 or less, 30 or less, 25 or less, 20 or less, 15 or less It is.

  In (I), one or several amino acids in the amino acid sequence of SEQ ID NO: 2 comprising 10 or more consecutive amino acid residues in the amino acid sequence deleted, added, substituted or inserted, and suffering from metabolic syndrome The polypeptide may be any polypeptide that can be used as a marker for risk determination, and in the amino acid sequence of SEQ ID NO: 2, 10 or more consecutive amino acid sequences in which one or several amino acids are deleted, added, substituted, or inserted. A polypeptide that consists of amino acid residues and can be used as a marker for risk assessment of metabolic syndrome is preferable.

  (J) is a polypeptide comprising 10 or more consecutive amino acid residues in the amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome A polypeptide consisting of 10 or more consecutive amino acid residues in the amino acid sequence having 90% or more homology with the amino acid sequence of SEQ ID NO: 2, which can be used as a marker for determining the risk of developing metabolic syndrome It is preferable that

  The mutation of one or several amino acid residues in (I) is the same as that described for (B) in item (1-1) above.

  The homology in (J) is the same as that described for (C) in item (1-1) above.

  Peptides selected from the group consisting of (F) to (J) include (F-2) to (F-7), (G-2) to (G-7) and (H-2) to (H-). A peptide selected from the group consisting of 7) is preferred, a peptide of (F-6) or (F-7) is more preferred, and a peptide of (F-7) is still more preferred.

(2) Antibody or Aptamer of the Present Invention The antibody or aptamer of the present invention is an antibody or aptamer that binds to the above-described marker peptide for risk assessment of metabolic syndrome of the present invention.

  By using the antibody or aptamer of the present invention, the amount or presence of the marker syndrome for determining metabolic syndrome morbidity of the present invention can be measured, and the risk of morbidity of metabolic syndrome can be determined. That is, when a biological sample collected from a subject is allowed to act on the antibody or aptamer of the present invention, the amount of the marker peptide bound to the antibody or aptamer is the amount of the biological sample collected from a healthy subject. When the amount of the marker peptide bound to the antibody or aptamer is high when acting on the aptamer, it is determined that the risk of suffering from metabolic syndrome is high.

  The antibody or aptamer can be prepared by a conventional method.

  An example of measuring the amount or presence of a marker peptide using the antibody or aptamer of the present invention is shown below. First, an antibody or aptamer is adsorbed on a carrier such as a microtiter plate by a known method such as physical adsorption or covalent bonding using a functional group, and then a biological sample is added after dilution as necessary and incubated. Next, a secondary antibody conjugated with a fluorescent substance, a chemiluminescent substance or an enzyme is added and incubated. For detection, evaluation is performed by adding each substrate and then measuring visible light by fluorescent or chemiluminescent substances or enzymatic reactions.

  The marker syndrome for determining metabolic syndrome risk of the present invention is a peptide present at a detectable level in a biological sample collected from a subject having a risk of suffering from metabolic syndrome. Therefore, the risk of suffering from metabolic syndrome can be determined using the presence of the marker peptide of the present invention as an index.

(3) Microarray of the present invention The microarray of the present invention is a microarray on which an antibody or aptamer that binds to the marker syndrome for determining metabolic syndrome morbidity described in the present invention is immobilized.

  A microarray is a generic term for devices in which substances that can bind to a substance to be measured are aligned and fixed on a carrier (base material). The material of the microarray carrier may be an inorganic material such as glass or an organic material such as nitrocellulose. The shape of the microarray carrier may be a membrane, a bead, or a plate.

  The microarray of the present invention can be produced by immobilizing the antibody or aptamer described in item (2) above on a carrier. For immobilization, devices such as a microarrayer and a spotter can be used.

  By using the microarray of the present invention, it is possible to measure the amount or the presence or absence of the metabolic syndrome morbidity risk marker peptide of the present invention, and it is possible to determine the metabolic syndrome morbidity risk. That is, when a biological sample collected from a subject is allowed to act on the microarray of the present invention, the amount of the marker peptide bound to the antibody or aptamer on the microarray corresponds to the biological sample collected from a healthy subject. When the amount of the marker peptide bound to the antibody or aptamer on the microarray is high, it is determined that the risk of suffering from metabolic syndrome is high.

  An example of measuring the amount of marker peptide using the microarray of the present invention is shown below. First, a biological sample is added to an immobilized antibody or aptamer on a microarray, a marker peptide in the biological sample is bound, and then a fluorescent substance, a chemiluminescent substance, or an enzyme is bound. Add antibody and incubate. Detection may be performed by adding each substrate and then measuring the fluorescent or chemiluminescent substance or visible light from the enzyme reaction.

(4) Determination Method of the Present Invention The determination method for the risk of developing metabolic syndrome according to the present invention measures the amount or presence of the marker peptide for determining the risk of metabolic syndrome according to claim 1 in a biological sample collected from a subject. Is the method.

  In determining the risk of suffering from metabolic syndrome from the amount of marker syndrome for risk assessment of metabolic syndrome, usually, comparison with a reference value is performed. As a reference value, for example, the amount of the marker peptide in a biological sample collected from a healthy person (preferably confirmed to be a healthy person by means other than the determination method of the present invention in advance), metabolic The amount of the marker peptide in a biological sample collected from a syndrome patient (preferably confirmed to be a metabolic syndrome patient in advance by means other than the determination method of the present invention) is preferable. The former is preferred.

  When the reference value is the value of a healthy person, the amount of the marker peptide for determining metabolic syndrome morbidity in the biological sample collected from the subject is higher than the amount of the marker peptide in the biological sample collected from the healthy person It is determined that the risk of developing metabolic syndrome is high.

  When the reference value is a value of a patient with metabolic syndrome, the amount of the marker peptide for risk assessment of metabolic syndrome in the biological sample collected from the subject is compared with the amount of the marker peptide in the biological sample collected from the patient with metabolic syndrome. If they are the same or high, it is determined that the risk of developing metabolic syndrome is high.

  In the determination method of the present invention, if the amount of the marker syndrome for determining metabolic syndrome morbidity in a biological sample collected from a subject is higher than the amount in a biological sample collected from a healthy person, It is determined that the risk of developing syndrome is high. On the other hand, if the amount of the metabolic syndrome disease risk marker peptide in the biological sample collected from the subject is the same or lower than the amount in the biological sample collected from healthy subjects, the risk of metabolic syndrome disease is Judge as low.

  Examples of biological samples include body fluids such as saliva, blood (whole blood, plasma, serum, etc.), urine, and tears. Of these, non-invasive and always collectable biological samples are preferable, and saliva is more preferable. Saliva is also suitable as a sample used for judgment at home. Saliva includes non-stimulated saliva and stimulated saliva, but stimulated saliva is preferred. Stimulated saliva can be easily collected by chewing paraffin gum.

  In the determination method of the present invention, the amount of marker syndrome for determining metabolic syndrome is determined by, for example, immunoassay using the antibody or aptamer, immunoassay using the microarray, mass spectrometry, RIA (radioimmunoassay). , ELISA (enzyme-linked immunosorbent assay), ECLIA (electrochemiluminescence immunoassay), and the like. The immunoassay using an antibody or aptamer is as already described in item (2). The immunoassay method using a microarray is as already described in item (3).

  Various types of mass spectrometers can be used for measurement by mass spectrometry. Examples of the mass spectrometer include GC-MS, LC-MS, FAB-MS, EI-MS, CI-MS, FD-MS, MALDI-MS, ESI-MS, HPLC-MS, FT-ICR-MS, CE-MS, ICP-MS, Py-MS, TOF-MS and the like can be mentioned, and any of these can be used.

  In the determination, multivariate analysis using the amount of two or more metabolic syndrome morbidity risk determination marker peptides as variables may be performed. Examples of multivariate analysis include logistic regression analysis, multiple regression analysis, principal component analysis, independent component analysis, factor analysis, discriminant analysis, quantification theory, cluster analysis, conjoint analysis, and multidimensional scaling method (MDS). Of these, logistic regression analysis is preferred.

  The marker peptide used in the determination method of the present invention may be one or more and may be a combination of two or more. When performing multivariate analysis as described above, two or more combinations are used. In the case of two or more, for example, there may be two, three, four, five, or six.

Examples of preferable combinations of marker peptides include the following combinations (1) to (3).
Combination (1): one or more peptides selected from the group consisting of (A) to (E) and one or more peptides selected from the group consisting of (F) to (J);
Combination (2): two or more peptides selected from the group consisting of (A) to (E); and Combination (3): two or more peptides selected from the group consisting of (F) to (J).

  The peptide selected from the group consisting of (A) to (E) and the peptide selected from the group consisting of (F) to (J) are as described in item (1).

Preferable examples of the combination (1) include the following combinations:
Combination (1-1): One or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (F-2) to (F-7), (G-2) to (G-7) and (H-2) to (H-7) A combination of
Combination (1-2): Combination of (A-6) and / or (A-7) with (F-6) and / or (F-7) Combination (1-3); (A-7) And (F-7) in combination Combination (1-4): (A-20) to (A-25), (B-20) to (B-25) and (C-20) to (C-25) ) And one or more peptides selected from the group consisting of: (F-2) to (F-7), (G-2) to (G-7) and (H-2) to (H-7) A combination with one or more peptides selected from the group;
Combination (1-5): A combination of (A-24) and / or (A-25) and (F-6) and / or (F-7);
Combination (1-6): Combination of (A-25) and (F-7) Combination (1-7): (A-2) to (A-7), (B-2) to (B-7) And one or more peptides selected from the group consisting of (C-2) to (C-7), and (A-14) to (A-19), (B-14) to (B-19) and ( One or more peptides selected from the group consisting of (C-14) to (C-19), (F-2) to (F-7), (G-2) to (G-7) and (H-2) ) To (H-7) in combination with one or more peptides selected from the group consisting of:
Combination (1-8): (A-6) and / or (A-7), (A-18) and / or (A-19), (F-6) and / or (F-7) In combination with;
Combination (1-9): Combination of (A-7), (A-19) and (F-7);
Combination (1-10): one or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (A-20) to (A-25), (B-20) to (B-25) and (C-20) to (C-25) A combination with one or more peptides selected from the group consisting of (F-2) to (F-7), (G-2) to (G-7) and (H-2) to (H-7);
Combination (1-11): (A-6) and / or (A-7), (A-24) and / or (A-25), (F-6) and / or (F-7) In combination with;
Combination (1-12): Combination of (A-7), (A-25) and (F-7);
Combination (1-13): one or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (A-14) to (A-19), (B-14) to (B-19) and (C-14) to (C-19) One or more peptides selected from the group consisting of: (A-20) to (A-25), (B-20) to (B-25) and (C-20) to (C-25); -2) to (F-7), (G-2) to (G-7) and a combination with one or more peptides selected from the group consisting of (H-2) to (H-7);
Combination (1-14): (A-6) and / or (A-7), (A-18) and / or (A-19), (A-24) and / or (A-25) And a combination of (F-6) and / or (F-7);
Combination (1-15): Combination of (A-7), (A-19), (A-25) and (F-7);
Combination (1-16): one or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (A-8) to (A-13), (B-8) to (B-13) and (C-8) to (C-13) One or more peptides selected from the group consisting of: (A-14) to (A-19), (B-14) to (B-19) and (C-14) to (C-19); -20) to (A-25), (B-20) to (B-25) and one or more peptides selected from the group consisting of (C-20) to (C-25), and (A-26) One or more peptides selected from the group consisting of (A-31), (B-26) to (B-31) and (C-26) to (C-31), and (F-2) to (F -7 , A combination of (G-2) ~ (G-7) and (H-2) ~ 1 or more peptides selected from (H-7) the group consisting of;
Combination (1-17): (A-6) and / or (A-7), (A-12) and / or (A-13), (A-18) and / or (A-19) A combination of (A-24) and / or (A-25), (A-30) and / or (A-31), and (F-6) and / or (F-7);
Combination (1-18): A combination of (A-7), (A-13), (A-19), (A-25), (A-31), and (F-7).

Examples of the combination (2) include the following combinations:
Combination (2-1): one or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (A-26) to (A-31), (B-26) to (B-31) and (C-26) to (C-31) A combination of
Combination (2-2): A combination of (A-6) and / or (A-7) and (A-30) and / or (A-31);
Combination (2-3): A combination of (A-7) and (A-31).
Combination (2-4): one or more selected from the group consisting of (A-2) to (A-7), (B-2) to (B-7) and (C-2) to (C-7) And one or more peptides selected from the group consisting of (A-14) to (A-19), (B-14) to (B-19) and (C-14) to (C-19) One or more peptides selected from the group consisting of: (A-20) to (A-25), (B-20) to (B-25) and (C-20) to (C-25); -26) to (A-31), (B-26) to (B-31) and a combination with one or more peptides selected from the group consisting of (C-26) to (C-31);
Combination (2-5): (A-6) and / or (A-7), (A-18) and / or (A-19), (A-24) and / or (A-25) And a combination of (A-30) and / or (A-31);
Combination (2-6): A combination of (A-7), (A-19), (A-25) and (A-31).

  Among the combinations (1-1) to (1-18) and (2-1) to (2-6), the combinations (1-2), (1-3), (1-5), (1-6) ), (1-8), (1-9), (1-11), (1-12), (1-14), (1-15), (1-17), (1-18), (2-2), (2-3), (2-5) and (2-6) are preferable, and combinations (1-3), (1-6), (1-9) and (1-12) , (1-15), (1-18), (2-3), and (2-6) are more preferable.

  The determination method of the present invention is suitable for evaluating a constitution related to the susceptibility of a subject to metabolic syndrome. Since the determination method of the present invention can evaluate the subject's risk of suffering from metabolic syndrome, it can also be used in preventive medicine. When a subject who is at risk of suffering from metabolic syndrome is treated or prevented by metabolic syndrome, the risk of suffering from metabolic syndrome proceeds in a decreasing direction, and the amount of marker peptide is also lowered accordingly. Therefore, evaluation of treatment or preventive measures can be performed by proceeding with the treatment or preventive measures and measuring the amount or presence of the marker peptide. Therefore, the biomarker of the present invention can be a biomarker for determining a therapeutic or prophylactic effect such as a drug administration effect. Similarly, the determination method of the present invention is also useful as a method for determining a therapeutic or prophylactic effect such as the administration effect of a drug of metabolic syndrome.

(5) Kit of the present invention The kit for determining the risk of suffering from metabolic syndrome of the present invention comprises the antibody or aptamer, or the microarray.

  The kit of the present invention preferably further contains a gum. This facilitates the collection of stimulated saliva as a biological sample. The gum may be any gum (paraffin gum or the like) that is normally used for collecting stimulated saliva.

Example 1
Comparison of expression levels of PB peptide fragment and GIPR peptide fragment in saliva <Evaluation method>
(1) Saliva specimen collection Stimulated saliva (saliva whose saliva secretion was promoted by chewing paraffin gum) was collected from subjects having the following characteristics.
A (healthy group): male, abdominal circumference less than 85 cm, BMI less than 25, none of the following items a) to d) (n = 10)
B (obese group): male, waist circumference 85 cm or more, or BMI 25 or more, and does not fall under any of the following items a) to d) (n = 10)
C (metabolic syndrome group): male, abdominal circumference of 85 cm or more or BMI of 25 or more, and corresponds to any one or more of the following items a) to d) in the blood test (n = 10)

Items a) Neutral fat: 150 mg / dL or more, or HDL-cholesterol: 40 mg / dL or less b) LDL-cholesterol: 140 mg / dL or more c) Fasting blood glucose: 110 mg / dL or more, or hemoglobin A1c: 5.8 % Or more d) Uric acid: 7.0 mg / dL or more

  In addition, although it is not a metabolic syndrome, the test subject who belongs to an obese group is estimated that the possibility of having a metabolic syndrome is high.

(2) Comprehensive analysis of saliva components by metabolome analysis The collected saliva was centrifuged to remove impurities, and the supernatant was subjected to LC-MS (Positive / Negative) and CE-MS (Anion / Cation). Salivary components were identified from the Rt value and Ms value.

<Evaluation results>
As a result of salivary metabolome analysis, the peak of the peptide ( ^PB peptide fragment (1) ^27-54 ) consisting of the amino acid sequence of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and the amino acid of SEQ ID NO: 2 It was recognized that the peak of the peptide consisting of the amino acid sequence of amino acid residues 264 to 275 in the sequence was significantly increased in the metabolic syndrome group. As a result of protein database search, the amino acid sequence of SEQ ID NO: 1 is the amino acid sequence of P-B peptide, the amino acid sequence of SEQ ID NO: 2 is the amino acid sequence of GIPR, and each of the above peptides is a fragment of each. did. The peak intensity of each subject group is shown in FIGS. Table 2 shows the significant difference test results for each group.

As shown in FIG. 1, Table 1 and Table 2, the peak intensity of the ^PB peptide fragment (1) ^27-54 is significantly increased in the obese group and the metabolic syndrome group as compared with the healthy group. Indicated. This means that when the peak intensity of P-B peptide fragment (1) ^27-54 of the subject exceeds 4.4, which is the peak intensity of the healthy subject group, it can be determined that the risk of suffering from metabolic syndrome is high. Is shown.

  On the other hand, as shown in FIG. 2, Table 1 and Table 2, the peak intensity of the GIPR peptide fragment was not significantly different in the obese group compared to the healthy group, but was significantly different in the metabolic syndrome group. It was shown to increase. This indicates that when the peak intensity of the GIPR peptide fragment of the subject exceeds 98.7, which is the peak intensity of the healthy group, it can be determined that the risk of suffering from metabolic syndrome is high.

Example 2
Determination of metabolic syndrome by measuring the amount of P-B peptide fragment in saliva Saliva was collected from 10 subjects corresponding to the metabolic syndrome group using the same criteria as in Example 1 in the same manner as in Example 1. -B peptide fragment (1) The peak intensity of ^27-54 was measured. As a result, in 7 of 10 saliva, the peak intensity of ^PB peptide fragment (1) ^27-54 exceeded 4.4, and the sensitivity of metabolic syndrome determination was calculated to be 70%. This result shows that the present invention is useful as a preliminary determination method of metabolic syndrome.

Example 3
Determination of metabolic syndrome by measuring the amount of GIPR peptide fragment in saliva Saliva was collected from 10 subjects corresponding to the metabolic syndrome group according to the same criteria as in Example 1 in the same manner as in Example 1, and the GIPR fragment was analyzed. Peak intensity was measured. As a result, in 9 of 10 saliva, the peak intensity of the peptide of SEQ ID NO: 2 exceeded 98.7, and the sensitivity of metabolic syndrome determination was calculated to be 90%. This result shows that the present invention is useful as a preliminary determination method of metabolic syndrome.

As described above, metabolomic analysis was performed on 40 saliva samples composed of 10 healthy persons, 10 obese persons, and 20 metabolic syndrome prone persons (10 subjects in Example 1 and 10 subjects in Examples 2 and 3). As a result, the amounts of P-B peptide fragment (1) ^27-54 and GIPR fragment were significantly increased in the metabolic syndrome prone group.

  The above results show that according to the present invention, the risk of suffering from metabolic syndrome can be easily determined with a certain degree of accuracy, and is useful as a preliminary determination method for risk of suffering from metabolic syndrome.

Example 4
Determination of metabolic syndrome by measuring amount of P-B peptide fragment in saliva <Evaluation method>
10 subjects corresponding to the healthy group, 10 corresponding to the obese group, and 20 subjects corresponding to the metabolic syndrome group based on the same criteria as in Example 1 (common with 20 subjects in Examples 1 to 3) ) 40 saliva samples were collected in the same manner as in Example 1. For these saliva samples, saliva metabolome analysis was performed in the same manner as in Example 1.

<Evaluation results>
As a result of salivary metabolomic analysis, the following four peptide fragment peaks were found to increase significantly in the metabolic syndrome group. The peak intensity of each subject group is shown in FIGS.

P-B peptide fragment (2) ^23-54 : peptide consisting of amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1 PB peptide fragment (3) ^55-79 : SEQ ID NO: Peptide / ^PB peptide fragment (4) ^consisting of amino acid sequences of amino acid residues 55 to 79 in one amino acid sequence (4) ^23-79 : of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1 Peptide / PB peptide fragment consisting of amino acid sequence (5) ^23-35 : Peptide consisting of amino acid sequence of amino acid residues ^{23 to 35} of amino acid sequence of SEQ ID NO: 1

As shown in FIG. 3 and Table 3, it was shown that the peak intensity of the ^PB peptide fragment (2) ^23-54 increases as compared with the healthy ^subject group. This means that when the peak intensity of the ^PB peptide fragment (2) ^23-54 of the subject exceeds 0.0, which is the peak intensity of the healthy subject group, it can be determined that the risk of suffering from metabolic syndrome is high. Is shown.

As shown in FIG. 4 and Table 3, the peak intensity of the ^PB peptide fragment (3) ^55-79 was shown to increase as compared with the healthy subject group. This means that when the peak intensity of P-B peptide fragment (3) ^55-79 of the subject exceeds 228.2 which is the peak intensity of the healthy subject group, it can be determined that the risk of suffering from metabolic syndrome is high. Is shown.

As shown in FIG. 5 and Table 3, the peak intensity of the ^PB peptide fragment (4) ^23-79 was shown to increase as compared with the healthy ^subject group. This means that when the peak intensity of the ^PB peptide fragment (4) ^23-79 of the subject exceeds 85.9, which is the peak intensity of the healthy subject group, it can be determined that the risk of suffering from metabolic syndrome is high. Is shown.

As shown in FIG. 6 and Table 3, the peak intensity of the ^PB peptide fragment (5) ^23-35 was shown to increase as compared with the healthy subject group. This means that when the peak intensity of the ^PB peptide fragment (5) ^{23-35 in} the subject exceeds 28.8 which is the peak intensity of the healthy group, it can be determined that the risk of suffering from metabolic syndrome is high. Is shown.

Example 5
Improvement of diagnostic accuracy by multivariate analysis (logistic regression analysis) of salivary components For the purpose of accurately determining metabolic syndrome for the five types of PB peptide fragments and GIPR fragments mentioned in Examples 1 to 4. Logistic regression analysis was performed as one of the variable analysis methods. Logistic regression analysis is an analysis technique that is generally used when predicting a qualitative variable (whether or not it is a metabolic syndrome in this application) using a plurality of objective variables (in this application, saliva components).

  Logistic regression analysis was performed on the saliva component combinations shown in Table 4 using the peak intensities of 10 healthy subjects and 20 metabolic syndrome groups collected in Examples 1 to 4. The obtained parameter estimation values (Table 5) were applied to Equation 1 to obtain a regression equation. The probability p of being healthy when the peak intensities of the group of 10 healthy subjects and the group of 20 metabolic syndrome collected in Examples 1 to 3 were again applied to this regression equation was calculated. If p> 0.5, it was judged as healthy, and if p <0.5, it was judged as metabolic syndrome, and the ratio that coincided with the actual judgment result was determined as the correct diagnosis rate (Table 6). As a result, it was suggested that diagnosis can be performed with high accuracy by using the combination of saliva components shown in Table 4.

Explanation of Formula 1 x1: ^PB peptide fragment (1) ^27-54 peak intensity x2: ^PB peptide fragment (2) ^23-54 peak intensity x3: ^PB peptide fragment (3) ^55-79 Peak intensity x4: ^PB peptide fragment (4) ^23-79 peak intensity x5: ^PB peptide fragment (5) ^23-35 peak intensity x6: GIPR fragment peak intensity

  In order to compare the diagnostic ability in the case of using a combination of these saliva components and the case of using a single saliva component listed in Examples 1 to 3, the area under the ROC (Receiver Operating Characteristic) curve was calculated. A plurality of cut-off values for classifying healthy subjects and metabolic syndrome subjects were set, and the sensitivity and specificity of each were calculated. Sensitivity is plotted on the vertical axis and 1-specificity is plotted on the horizontal axis, and an ROC curve is drawn. The larger the area under the ROC curve, the higher the diagnostic ability (written by Toshio Mori, “Easy to understand medical statistics”, Medical Tribune, Inc., p254). As a result, as shown in Table 7, it was suggested that the diagnostic ability is enhanced by the combination of saliva components as compared with the case using a single saliva component.

Claims (11)

The following (A-7), (A-13), (A-19), (A-25), (A-31), (B-7), (B-13), (B-19), (B-25), (B-31), (C-7), (C-13), (C-19), (C-25), (C-31), (F-7), (G -7), an antibody or aptamer that specifically binds to a marker for determining risk of developing metabolic syndrome, which is a peptide selected from the group consisting of (H-7).
(A-7) a polypeptide comprising amino acid residues Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-13) a polypeptide comprising amino acid residues Nos. 23 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-19) a polypeptide comprising amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-25) a polypeptide consisting of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-31) a polypeptide comprising amino acid residues Nos. 23 to 35 in the amino acid sequence of SEQ ID No. 1;
(B-7) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 27 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-13) Determination of risk of suffering from metabolic syndrome consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-19) Determination of risk of developing metabolic syndrome, comprising an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-25) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-31) An amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determining the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(C-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-13) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-19) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-25) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue of amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-31) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 23 to 35 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(F-7) a polypeptide comprising amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2;
(G-7) The amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 consists of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determines the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(H-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome The following (A-7), (A-13), (A-19), (A-25), (A-31), (B-7), (B-13), (B-19), (B-25), (B-31), (C-7), (C-13), (C-19), (C-25), (C-31), (F-7), (G -7) A microarray in which an antibody or aptamer that specifically binds to a metabolic syndrome disease risk determination marker, which is a peptide selected from the group consisting of (H-7), is immobilized on a carrier.
(A-7) a polypeptide comprising amino acid residues Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-13) a polypeptide comprising amino acid residues Nos. 23 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-19) a polypeptide comprising amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-25) a polypeptide consisting of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-31) a polypeptide comprising amino acid residues Nos. 23 to 35 in the amino acid sequence of SEQ ID No. 1;
(B-7) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 27 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-13) Determination of risk of suffering from metabolic syndrome consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-19) Determination of risk of developing metabolic syndrome, comprising an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-25) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-31) An amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determining the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(C-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-13) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-19) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-25) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue of amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-31) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 23 to 35 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(F-7) a polypeptide comprising amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2;
(G-7) The amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 consists of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determines the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(H-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome In the biological sample collected from the subject, the following (A-7), (A-13), (A-19), (A-25), (A-31), (B-7), (B- 13), (B-19), (B-25), (B-31), (C-7), (C-13), (C-19), (C-25), (C-31) , (F-7), (G-7), (H-7), a method for determining the risk of metabolic syndrome, comprising measuring the amount or presence of a marker for determining the risk of metabolic syndrome, which is a peptide selected from the group consisting of .
(A-7) a polypeptide comprising amino acid residues Nos. 27 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-13) a polypeptide comprising amino acid residues Nos. 23 to 54 in the amino acid sequence of SEQ ID No. 1;
(A-19) a polypeptide comprising amino acid residues 55 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-25) a polypeptide consisting of amino acid residues 23 to 79 in the amino acid sequence of SEQ ID NO: 1;
(A-31) a polypeptide comprising amino acid residues Nos. 23 to 35 in the amino acid sequence of SEQ ID No. 1;
(B-7) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 27 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-13) Determination of risk of suffering from metabolic syndrome consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-19) Determination of risk of developing metabolic syndrome, comprising an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residue positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-25) Determination of risk of developing metabolic syndrome, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted in the amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1. A polypeptide that can be used as a marker for an animal;
(B-31) An amino acid sequence of amino acid residues 23 to 35 of the amino acid sequence of SEQ ID NO: 1, consisting of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determining the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(C-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues of amino acid residues 27 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-13) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residues 23 to 54 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-19) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 55 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(C-25) A polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue of amino acid residues 23 to 79 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for determining the risk of developing metabolic syndrome ;
(C-31) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid residue at positions 23 to 35 of the amino acid sequence of SEQ ID NO: 1 and can be used as a marker for risk assessment of metabolic syndrome ;
(F-7) a polypeptide comprising amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2;
(G-7) The amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 consists of an amino acid sequence in which one amino acid is deleted, added, substituted or inserted, and determines the risk of suffering from metabolic syndrome A polypeptide that can be used as a marker for an animal;
(H-7) a polypeptide comprising an amino acid sequence having 90% or more identity with the amino acid sequence of amino acid residues 264 to 275 of the amino acid sequence of SEQ ID NO: 2 and can be used as a marker for risk assessment of metabolic syndrome Claims that the risk of developing metabolic syndrome is high when the amount of the marker for risk assessment of metabolic syndrome in a biological sample collected from a subject is higher than the amount in the biological sample collected from a healthy subject. 3. The determination method according to 3 . Determining in a biological sample collected from a subject, the amount of the metabolic syndrome morbidity risk marker for determination is, if the same or higher than the amount in a biological sample taken from patients with metabolic syndrome, and high metabolic syndrome risk of acquiring The determination method according to claim 3 . Determining the amount of the metabolic syndrome morbidity risk marker for determination, in the mass spectrometry, and immunoassay using an antibody or aptamer according to claim 1, the immunoassay using the microarray of claim 2 carried out by a method selected from the determination method according to any one of claims 3-5. The determination method according to claim 3 , wherein multivariate analysis is performed using a combination of two or more markers for determining the risk of developing metabolic syndrome as variables. The determination method according to claim 7 , wherein the multivariate analysis is logistic regression analysis. Biological sample, the determination method according to any one of claims 3-8 is saliva. Antibodies or aptamers according to claim 1, or comprises a microarray of claim 2, metabolic syndrome morbidity risk determination kit. The kit according to claim 10 , further comprising a gum.